The aim of this study was to investigate the effect of sorafenib combined with daunorubicin on leukemic k562 cell line. The inhibitory effect of sorafenib alone and its combination with daunorubicin on K562 cell proliferation was detected by MTT method; the synergistic effect was measured by CDI (coefficient of drug interaction); the apoptosis of K562 cells was observed by flow cytometry with Hoechst 33258 staining. The results showed that the sorafenib alone or its combination with daunorubicin could significantly inhibit K562 cell proliferation and the combination of both drugs displayed synergistic effect on K562 cells, meanwhile the apoptotic cells increased. It is concluded that the combination of sorafenib and daunorubicin has a obviously synergistic inhibitory effect on leukemic cell line K562.
Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Daunorubicin ; pharmacology ; Drug Synergism ; Humans ; K562 Cells ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology

To investigate the effects of sorafenib and octreotide combination treatment on cellular proliferation and explore the underlying molecular mechanisms by using an in vitro cell culture system with the human hepatoma cell line, HepG2. HepG2 cells were treated with different concentrations of sorafenib and octreotide alone or in combination. Untreated HepG2 cells were used as controls. Treatment-induced cytotoxicity was determined with the cell counting kit-8 by Sigma-Aldrich, and rate of apoptosis was detected by flow cytometry. Fluorescent microscopy was used to observe rates of cell growth under the various treatments. Treatment-induced changes in protein expressions were detected by enzyme-linked immunosorbent assay (for vascular endothelial growth factor (VEGF)) and Western blotting (for the Mcl-1 apoptosis mediator and the ERK1/2 and PERK1/2 kinases). Sorafenib and octreotide, used alone or in combination, inhibited proliferation and induced apoptosis in HepG2 cells. Combination treatment was more effective than either mono-treatment (F = 200.398, P less than 0.05). Fluorescent microscopy showed that combination treatment stimulated phosphatidylserine, the marker of early apoptosis, better than either mono-treatment. VEGF expression in cultures exposed to combination treatment was also significantly lower than in mono-treatment or untreated control cultures (F = 1019.725, P less than 0.05). Western blotting showed that octreotide mono-treatment had no effect on Mcl-1 expression (vs. control group; P more than 0.05) and that combination treatment significantly lowered Mcl-1 expression (vs. mono-treatment and control groups; P less than 0.05). None of the treatments affected ERK1/2 expression (all, P more than 0.05), while all treatments significantly lowered PERK1/2 expression (vs. control group; F = 2.401, P less than 0.05) and the combination treatment lowered PERK1/2 significantly more than either mono-treatment (P less than 0.05). Sorafenib and octreotide can inhibit proliferation and induce apoptosis in the human hepatoma cell line, HepG2. Combination treatment is significantly more efficacious (P less than 0.05) and produced synergistic effects. The mechanism underlying this phenomenon may depend on synergistic inhibition of VEGF, the anti-apoptotic protein Mcl-1, and the proliferation-inducing PERK1/2.
Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Cell Proliferation ; drug effects ; Hep G2 Cells ; drug effects ; Humans ; Niacinamide ; analogs & derivatives ; Octreotide ; pharmacology ; Phenylurea Compounds ; Pyridines ; pharmacology

BACKGROUNDKeratinocytes play a crucial role in the biological function of skin barrier. The relationship between sodium lauryl sulfate (SLS) and keratinocytes has been studied. However, the cytotoxicity and effects of sodium dodecyl benzene sulfonate (SDBS), a common detergent similar to SLS, on keratinocytes are still not known. This study aimed to investigate the effects of SDBS on cytotoxicity and expression of proinflammatory cytokines in cultured human keratinocytes.

METHODSThis study was carried out using the keratinocytes cell line, HaCaT cells. The cytotoxicity of SDBS on HaCaT cells was evaluated with cell counting kit-8 (CCK-8) and phase-contrast microscopy. After exposure to different concentrations of SDBS, the total RNA of the HaCaT cells was extracted for evaluating the relative mRNA expression of IL-1α, IL-6, IL-8, and TNF-α by qPCR. The supernatants of cells were collected for measuring the levels of IL-6 and IL-8 by enzyme-linked immunosorbent assay (ELISA).

RESULTSSDBS at concentrations of 20 µg/ml and over showed direct cytotoxicity and induced morphological changes of the HaCaT cells. The mRNA expressions of IL-1α, IL-6, IL-8, and TNF-a in different concentrations of SDBS at different time were comparable with that of controls. SDBS at concentrations of 5, 10, and 15 µg/ml had no significant effects on IL-6 and IL-8 excretion from HaCaT cells after 24-hour exposure. Moreover, no significant effects on the IL-6 and IL-8 excretion were found after 10 and 15 µg/ml SDBS stimulations for 6, 12, and 24 hours, respectively.

CONCLUSIONSDBS at higher concentrations had cytotoxicity on HaCaT cells but had no effects on the mRNA expression of IL-1α, IL-6, IL-8, and TNF-a, that was different from SLS.

Benzenesulfonates ; pharmacology ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-1alpha ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Keratinocytes ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

This study was aimed to investigate the effect of multikinase inhibitor sorafenib on the proliferation and apoptosis of U937 cells and its possible mechanism. U937 cells were treated with different concentrations of sorafenib for 48 hours. Cell viability was determined by Cell Counting Kit-8; cell apoptosis and cell ratio in cell cycle were detected by flow cytometry with Annexin V/PI staining and PI staining respectively; expressions of GSK-3β, β-catenin and cyclin-D1 were assayed by Western blot. The results showed that the proliferation of U937 cells was inhibited by sorafenib in a dose-dependent manner (p < 0.05). Sorafenib induced cell apoptosis and cell cycle G(1)/G(0) arrest. Compared with results of Western blot before treatment, expression of inactivated GSK-3β, β-catenin and Cyclin-D1 down-regulated in a dose-dependent manner after treatment with sorafenib, this same changes were observed after up-regulation of inactivated GSK-3β by LiCl (p < 0.05). It is concluded that sorafenib inhibits the proliferation of U937 cells and induces cell apoptosis through reducing negative regulation of WNT signal pathway on inactivated GSK-3β and down-regulating β-catenin and cyclin-D1 level, which result in U937 cell cycle G(1)/G(0) arrest.
Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology ; U937 Cells ; Wnt Signaling Pathway ; beta Catenin ; metabolism

Epidermal growth factor receptor (EGFR)-targeted therapies have been effective in some cancers, but not in hepatocellular carcinoma (HCC). The aim of this study was to investigate the drug potential to overcome multi-drug resistance in HCC cells. Thirteen drug-sensitive HCC cells were assessed using the CCK-8 assay. G0-G1 arrest was measured by FACS. Western blot analysis was used to detect the key enzymes in both the Ras/Raf and PI3K pathways. When establishing the IC50 of HCC to several drugs, including EKB-569, sorafenib, erlotinib, gefitinib, pazopanib, and brivanib, SK-Hep1 cells treated with EKB-569 have shown the highest (72.8%-86.4%) G0-G1 arrest and decreased the phosphorylation of AKT and ERK at the protein level. We found that EKB-569 had higher efficacy in HCC, compared to first generation, reversible EGFR-TK inhibitors. Furthermore, the combination of sorafenib and EKB-569 showed a synergistic effect to inhibit proliferation of SNU-475, previously the most resistant cell to EGFR-TKIs. Therefore, novel EKB-569 in combination with sorafenib may be able to overcome HCC resistance to EGFR-TK inhibitors.
Aminoquinolines/*pharmacology ; Aniline Compounds/*pharmacology ; Antineoplastic Agents/*pharmacology ; Antineoplastic Combined Chemotherapy Protocols/pharmacology ; Benzenesulfonates/pharmacology ; Carcinoma, Hepatocellular/*drug therapy/pathology ; Cell Cycle Checkpoints/drug effects ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; Liver Neoplasms/drug therapy/pathology ; Mitogen-Activated Protein Kinases/*metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt/*metabolism ; Pyridines/pharmacology ; Receptor, Epidermal Growth Factor/*antagonists & inhibitors

OBJECTIVETo investigate the inhibitory effect of sorafenib in combination with arsenic trioxide (As2O3) on hepatocellular carcinoma cells and explore the mechanisms of the synergetic antitumor effects of the two agents.

METHODSHepG2 cells were treated with sorafenibor, As2O3 alone or their combination, with the untreated cells used as the control. The inhibitory effect of the treatment was analyzed by MTT assay, and the cell apoptosis and mitochondrial transmembrane potential (delta phi m) were detected by flow cytometry. Western blotting was performed to examine the expressions of ERK and pERK in the cells.

RESULTSSorafenib and As2O3 used alone or in combination both inhibited the proliferation of HepG2 cells, and a synergistic effect of the two agents was noted in their combined action (P<0.05). Combined treatment of the cells resulted in significantly higher apoptsis rate than that in the other groups (P<0.05), and was associated with a more obvious decrease in delta phi m. The expression of ERK was not affected by the two agents used either alone or in combination, but pERK expression was significantly lowered in the cells after combined treatment for 24 h.

CONCLUSIONA synergistic effect is observed between the sorafenib and As2O3 in their inhibition of HepG2 cell proliferation, the mechanisms of which may involve reduction of mitochondrial transmembrane potential and Raf/MEK/ERK pathway inhibition.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Benzenesulfonates ; pharmacology ; Blotting, Western ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Drug Synergism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Flow Cytometry ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Niacinamide ; analogs & derivatives ; Oxides ; pharmacology ; Phenylurea Compounds ; Pyridines ; pharmacology

OBJECTIVETo investigate the inhibitory effect of sorafenib in combination with cisplatin (DDP) on the proliferation of hepatocellular carcinoma cells and explore the molecular mechanisms.

METHODSThe inhibitory effect of sorafenib and DDP treatment on HepG2 cell proliferation in vitro was assessed by MTT assay. The cell cycle changes and the apoptotic rate of the treated cells were detected by flow cytometry, and the expressions of ERK and pERK examined using Western blotting.

RESULTSSorafenib and DDP alone both significantly inhibited the proliferation of HepG2 cells, showing a synergistic effect of their actions in combined use (P<0.05). Sorafenib and DDP alone caused cell cycle arrest at G(1) and G(2) phases, respectively. Combined use of the two drugs resulted in significant reduction of the S-phase cell percentage and cell cycle arrest at G(1) and G(2) phases. The coadministration of the drugs significantly increased the apoptosis rate of the cells as compared with the that of the cells with sorafenib or DDP treatment alone (P<0.05). Sorafenib and DDP, used alone or in combination, did not produce obvious effect on ERK expression. Sorafenib treatment for 24 h reduced pERK expression in the HepG2 cells, and the effect was enhanced by combined treatment with sorafenib and DDP.

CONCLUSIONSSorafenib and DDP show a synergistic effect in inhibiting the proliferation and inducing apoptosis of HepG2 cells. The mechanisms of this synergistic effect can be closely related to the double blockage of the cell cycle and Raf/MEK/ERK pathway inhibition.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Blotting, Western ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Drug Synergism ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Flow Cytometry ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology

OBJECTIVETo explore the role of sorafenib in reversing multidrug resistance (MDR) in hepatoma BEL-7402/FU cells and its possible mechanisms.

METHODSMTT colorimetric assay was used to obtain the dose-response curve of sorafenib in BEL-7402/FU cells, and flow cytometry performed to assess the effect of sorafenib on Rho123 concentration in the cells. The optimal dose of sorafenib for cell treatment was determined according to the results of MTT assay and flow cytometry. MTT assay was employed to evaluate the effect of sorfenib on the cytotoxicity of the antitumor drugs, flow cytometry performed to determine the expression of cell membrane transport protein (P-gp), and RT-PCR used to detect mdr1 gene expression in the cells treated with sorafenib at the optimal dose.

RESULTSSorafenib at the concentration of 4 micromol/L, efficiently reversed the MDR of the cells with minimal side effects. At the concentration of 4 micromol/L, sorafenib partially reversed the drug resistance of BEL-7402/FU cells to ADM, 5-FU, GEM and DDP, with reversal indexes of 2.98, 7.16, 1.99 and 10.08, respectively. Treatment of the cells with 4 micromol/L, sorafenib also partially down-regulated P-gp expression in BEL-7402/FU cells, and caused a reduction of mdr1 gene expression by 27.3% in comparison with the control cells.

CONCLUSIONSorafenib can reverse MDR in human hepatoma cells probably in association with down-regulation of mdr1 gene expression and increased accumulation of the chemotherapeutic agents in the cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Benzenesulfonates ; pharmacology ; Cell Line, Tumor ; Down-Regulation ; drug effects ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Liver Neoplasms ; genetics ; Niacinamide ; analogs & derivatives ; Phenylurea Compounds ; Pyridines ; pharmacology

Antineoplastic Agents ; pharmacology ; therapeutic use ; Benzenesulfonates ; pharmacology ; therapeutic use ; Carcinoma, Hepatocellular ; diagnosis ; surgery ; therapy ; Diagnosis, Differential ; Humans ; Liver Neoplasms ; diagnosis ; surgery ; therapy ; Liver Transplantation ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Niacinamide ; analogs & derivatives ; Oligonucleotide Array Sequence Analysis ; Phenylurea Compounds ; Polymerase Chain Reaction ; Pyridines ; pharmacology ; therapeutic use ; Retrospective Studies ; Survival Rate

OBJECTIVETo observe the signal transducers and activator of transcriptions (STATs) protein expression changes and investigate the functional role of STATs pathway in case of high glucose-induced cardiac fibroblasts (CFs) proliferation and collagen deposition in vitro.

METHODSRat cardiac fibroblasts were isolated from 1- to 3-day-old SD rats, cells from the second to fourth passages were used for the experiment. CFs were cultured in Dulbecco's modified Eagle's medium, supplemented with 5.5 mmol/L glucose (NG), 5.5 mmol/L glucose plus 19.4 mmol/L mannose (OC) or 25 mmol/L glucose (HG) in the presence of absence of STAT1 inhibitor (fludarabine, FLU) and STAT3 inhibitor (S3I-201). After 24 h and 48 h culture in vitro, the proliferation of CFs was measured by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. After 12 h and 24 h culture in vitro, the production of type I and III collagen was evaluated using real-time quantitative PCR and ELISA. After 0, 30, 60 and 120 min culture in vitro, the phosphorylated expression of STAT1 and STAT3 was analyzed by Western blot.

RESULTSCFs proliferation was significantly enhanced post 24 h and 48 h HG stimulation, and procollagen I and III mRNA expression was significantly upregulated post 12 h and 24 h HG stimulation. Deposition of collagen I and III was also significantly increased post 24 h and 72 h HG stimulation. STAT1 phosphorylation in CFs was increased after 120 min HG stimulation and STAT3 phosphorylation in CFs was increased post 60 min and 120 min HG stimulation. FLU and S3I-201 could inhibit HG-induced CFs proliferation and suppress of which was stimulated by FLU and S3I-201 could both suppress upregulated procollagen I and III mRNA expression and the deposition of collagen types I and III post HG stimulation. STAT1 phosphorylation inhibition resulted in less mRNA downregulation of procollagen type III than that of procollagen type I post 12 h HG stimulation. The STAT3 phosphorylation inhibition resulted in more significantly upregulated procollagen type III mRNA expression than procollagen type I mRNA expression at 12 h post HG stimulation.

CONCLUSIONHG could enhance the protein expression of phosphorylated STAT1 and STAT3 in CFs, which are responsible for HG-induced increased CFs proliferation and collagen deposition in vitro.

Aminosalicylic Acids ; pharmacology ; Animals ; Benzenesulfonates ; pharmacology ; Cell Proliferation ; Cells, Cultured ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Fibroblasts ; cytology ; drug effects ; Glucose ; adverse effects ; Myocardium ; cytology ; Phosphorylation ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; STAT1 Transcription Factor ; metabolism ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Up-Regulation ; Vidarabine ; analogs & derivatives ; pharmacology